Target (protein/gene name): Adenylate Kinase/Myokinase NCBI Gene # or RefSeq#:
Gene #: 5651684
RefSeq #: XM_001683025.1 Protein ID (NP or XP #) or Wolbachia#: XP_001683077.1 Organism (including strain): Leishmania major strain Friedlin
Etiologic Risk Group (see link below): The link on the site is broken. Disease Information
Leishmaniasis is a zoonosis skin infection that is transmitted through sand fly bites into humans, making it an endemic in tropical and subtropical parts of the world. Lesions form around source of infection and then ulcerate, possibly becoming secondarily infected with bacteria, which can be debilitating. Link to TDR Targets page (if present): TDR Target Page Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): NCBI Database
Essentiality of this protein: Shown to be essential in many orthologs, and inhibition leads to birth defects, infertility, sterility, and usually significant loss of fitness within 3 days.
Complex of proteins: Works with AMP in a multimer structure ATP + AMP <=> 2ADP
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): Druggability index of 0.8, according to TDRTargets.
Use of adenine nucleotide derivatives to assess the potential of exo-active-site-directed reagents as species- or isozyme-specific enzyme inactivators. 4. Interactions of adenosine 5'-triphosphate derivatives with adenylate kinases from Escherichia coli and rat tissues. (1982).[link]
Use of adenine nucleotide derivatives to assess the potential of exo-active-site-directed reagents as species- or isozyme-specific enzyme inactivators. 3. Synthesis of adenosine 5'-triphosphate derivatives with N6- or 8-substituents bearing iodoacetyl groups. (1982). [link]
Species- or isozyme-specific enzyme inhibitors. 4. Design of a two-site inhibitor of adenylate kinase with isozyme selectivity. (1982). [link]
Species- or isozyme-specific enzyme inhibitors. 7. Selective effects in inhibitions of rat adenylate kinase isozymes by adenosine 5'-phosphate derivatives. (1982). [link]
Species- or isozyme-specific enzyme inhibitors. 8. Synthesis of disubstituted two-substrate condensation products as inhibitors of rat adenylate kinases. (1982). [link]
Expression Information (has it been expressed in bacterial cells): Can be expressed in E.coli cells Purification Method
Affi-Gel Blue Gel column chromatography, Q-Sepharose column chromatography, and S-200 gel filtration [link]
Image of protein (PyMol with features delineated and shown separately):
Protein chains are colored from the N-terminal to the C-terminal using a rainbow (spectral) color gradient
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
mssndglsed tvmyikdnni gqlmeyilrc iitdkptkpl eyvheltasp lpprvvlagp pasgkgtqar hicsyykrai gkkpvhvssg dllraevaqg thlgkiaenf mqrgelvpds liisiirnrl tqedavmngw lldgfprtrs qaialdaagl cprifvvldt pddvlfgrve grrtdpvtgi iyhlkynppp endtallerl qhrdddtrev lgprletyhs mveglldyyg simyhvdgnr peaaitkdit eylqnhnva *length of your protein in Amino Acids: 269 aa Molecular Weight30.0602 kDa Extinction coefficient23380, assuming all Cys residues are reduced
Do Not Need this info for Spring (but still copy these lines to your Target page for now) Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers): (link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.
NCBI Gene # or RefSeq#:
Gene #: 5651684
RefSeq #: XM_001683025.1
Protein ID (NP or XP #) or Wolbachia#: XP_001683077.1
Organism (including strain): Leishmania major strain Friedlin
Etiologic Risk Group (see link below):
The link on the site is broken.
Disease Information
Leishmaniasis is a zoonosis skin infection that is transmitted through sand fly bites into humans, making it an endemic in tropical and subtropical parts of the world. Lesions form around source of infection and then ulcerate, possibly becoming secondarily infected with bacteria, which can be debilitating.
Link to TDR Targets page (if present): TDR Target Page
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): NCBI Database
Essentiality of this protein:
Shown to be essential in many orthologs, and inhibition leads to birth defects, infertility, sterility, and usually significant loss of fitness within 3 days.
Complex of proteins:
Works with AMP in a multimer structure
ATP + AMP <=> 2ADP
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
Druggability index of 0.8, according to TDRTargets.
*EC#: 2.7.4.3.
Link to BRENDA EC# page: BRENDA 2.7.4.3.
Enzyme Assay
Enzymatic Essay for Myokinase (2.7.4.3.) [Sigma Aldrich]
*(If supplier not named, Sigma Aldrich)
hexokinase ≥65 units/mg protein, glucose-6-phosphate dehydrogenase ≥75 units/mg protein (biuret)
Structure (PDB or Homology model)
PDB: 1Y63
Current Inhibitors:
- P1,P5-(bis adenosine)-5'-pentaphosphate [link]
Expression Information (has it been expressed in bacterial cells):Can be expressed in E.coli cells
Purification Method
Affi-Gel Blue Gel column chromatography, Q-Sepharose column chromatography, and S-200 gel filtration [link]
Image of protein (PyMol with features delineated and shown separately):
Protein chains are colored from the N-terminal to the C-terminal using a rainbow (spectral) color gradient
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
mssndglsed tvmyikdnni gqlmeyilrc iitdkptkpl eyvheltasp lpprvvlagp pasgkgtqar hicsyykrai gkkpvhvssg dllraevaqg thlgkiaenf mqrgelvpds liisiirnrl tqedavmngw lldgfprtrs qaialdaagl cprifvvldt pddvlfgrve grrtdpvtgi iyhlkynppp endtallerl qhrdddtrev lgprletyhs mveglldyyg simyhvdgnr peaaitkdit eylqnhnva
*length of your protein in Amino Acids: 269 aa
Molecular Weight30.0602 kDa
Extinction coefficient23380, assuming all Cys residues are reduced
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html).
Gene Sequence
1 atgtcctcca acgacggcct ctccgaggac acggtcatgt acatcaagga caacaacatt
61 gggcagctga tggagtacat cttgcgttgc atcatcaccg acaaaccaac caagccgctc
121 gaatatgtgc atgagttgac agcctcgccg ctgccgccgc gggtggtcct cgctgggccg
181 ccggcaagcg gcaagggcac gcaggcgcgc cacatctgct cctattacaa gcgcgcgatt
241 ggcaagaagc cagtgcatgt ctcatcgggc gatcttcttc gcgctgaagt ggctcagggc
301 acccacctgg gcaagattgc cgagaacttc atgcagcgcg gtgagctcgt gccggatagc
361 ctcatcatca gcattatccg caaccgactg acgcaggagg acgcggtgat gaacggatgg
421 ctgctggacg gcttcccgcg cacccgctct caagcgattg cgctggacgc cgcaggcctg
481 tgcccccgca tctttgtggt cctcgacacc ccagatgatg ttttgtttgg ccgcgtcgaa
541 ggccgtcgca ccgacccggt caccggcatc atctaccacc tcaagtacaa cccgccgccg
601 gagaacgaca cggcgctgct cgagcggctg cagcaccgcg acgatgacac ccgcgaggtg
661 cttggccctc gtctggagac ataccactca atggtcgagg gtctgctgga ctactacggg
721 tccatcatgt atcacgtcga cgggaaccga ccggaggcag ccatcacgaa ggacattacc
781 gagtacctcc aaaaccacaa tgtagcgtag
*GC% Content for gene: 60.99%
output
ATG TCT AGC AAC GAC GGC CTG TCG GAG GAC ACC GTC ATG TAC ATC AAG GAC AAT AAC ATT GGC CAG CTT ATG GAG TAC ATT CTC CGC TGC ATC ATC ACG GAT AAG CCG ACT AAG CCC CTC GAG TAC GTG CAT GAG CTC ACT GCG TCT CCG CTG CCA CCA CGG GTG GTC CTG GCC GGC CCG CCG GCC TCC GGC AAG GGT ACG CAG GCG CGC CAT ATT TGC TCG TAC TAC AAG CGC GCG ATC GGC AAG AAG CCG GTG CAC GTG TCC TCC GGC GAC CTG CTC CGC GCC GAG GTG GCC CAG GGT ACG CAC TTG GGT AAG ATT GCG GAG AAC TTC ATG CAG CGT GGC GAG CTC GTG CCC GAC AGC TTG ATT ATC TCC ATC ATC CGC AAC CGC CTG ACT CAG GAG GAC GCC GTG ATG AAT GGG TGG CTC CTG GAT GGT TTC CCG CGG ACG CGG TCG CAG GCC ATC GCC CTT GAT GCG GCC GGC CTC TGT CCG CGC ATC TTC GTC GTC TTG GAC ACC CCG GAC GAC GTG CTG TTT GGC CGG GTG GAG GGC CGC CGC ACT GAT CCG GTG ACG GGC ATT ATC TAC CAC CTC AAG TAC AAC CCG CCG CCG GAG AAT GAC ACC GCC CTG CTC GAG CGC CTG CAG CAT CGC GAC GAT GAT ACA CGC GAG GTC CTG GGT CCG CGC CTG GAG ACG TAC CAC AGC ATG GTG GAG GGC CTG CTG GAC TAT TAC GGT AGC ATC ATG TAC CAC GTG GAC GGC AAC CGG CCT GAG GCA GCG ATC ACG AAG GAC ATT ACA GAG TAC CTG CAG AAC CAC AAC GTG GCA TGA
*GC% Content for gene (codon optimized): 62.10%
Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.